Mitochondrial augmentation therapy

Abstract

The present invention provides methods and compositions which cause the bone marrow to increase production of leukocyte cells. Specifically, the invention provides methods and compositions for increasing levels of CD45+ cells in a subject by providing mitochondrially-enriched cells. Further the present application provides methods for increasing bone marrow cellularity, engraftment of CD34+ cells and differentiation.

1 . A method of increasing levels of leukocyte cells in a subject comprising:
 (a) obtaining target cells from a subject;   (b) obtaining exogenous mitochondria from a donor cell;   c) producing mitochondrially-enriched target cells by contacting the target cells with the exogenous mitochondria under conditions allowing the exogenous mitochondria to enter the target cells; and   (d) administering the mitochondrially-enriched target cells to the subject,   
       wherein the mitochondrial content of the mitochondrially-enriched target cells is detectably higher than the mitochondrial content of the target cells, thereby increasing the levels of leukocyte cells in a subject. 
     
     
         2 . The method of  claim 1 , wherein the target cells are selected from the group consisting of: pluripotent stem cells, embryonic stem cells, induced pluripotent stem cells, mesenchymal stem cells, hematopoietic stem cells, hematopoietic progenitor cells, common myeloid progenitor cells, common lymphoid progenitor cells, CD34+ cells, and any combination thereof. 
     
     
         3 . The method of  claim 1 , wherein the target cells are CD34+ cells. 
     
     
         4 . The method of  claim 1 , wherein the target cells are obtained from whole blood, blood fractions, peripheral blood, PBMC, serum, plasma, adipose tissue, placenta, oral mucosa, blood, umbilical cord blood or bone marrow. 
     
     
         5 . The method of  claim 1 , wherein the subject has a disease or disorder. 
     
     
         6 . The method of  claim 5 , wherein the disease or disorder is selected from the group consisting of: age related disorders, cancer, muscle diseases and disorders, glycogen-storage diseases and disorders, vascular endothelium disorder or diseases, brain disorder or brain disease, placental disorder or placental disease, thymus disorder or thymus disease, autoimmune diseases, renal disease or renal disorder, primary mitochondrial disease, pancreas disorder or pancreas disease, prostate disorder or prostate disease, kidney disorder or kidney disease, blood disorder or blood disease, heart disease or heart disorder, skin disorder or skin disease, immune and inflammatory diseases and disorders, bone disease or bone disorder, gastro-intestinal disease or gastro-intestinal disorder, eye disease or eye disorder and infection. 
     
     
         7 . The method of  claim 1 , wherein the exogenous mitochondria are isolated or partially purified frozen-thawed human mitochondria. 
     
     
         8 . The method of  claim 1 , wherein the exogenous mitochondria constitute at least 1% of the total mitochondria content in the mitochondrially-enriched target cells. 
     
     
         9 . The method of  claim 1 , wherein the exogenous mitochondria are derived from a human cell or tissue selected from the group consisting of placenta, placental cells grown in culture, blood cells, and stem cell. 
     
     
         10 . The method of  claim 1 , wherein the mitochondrial content of the mitochondrially-enriched target cells is determined by an assay selected from the group consisting of: content of at least one mitochondrial protein selected from SDHA and COX1; activity level of citrate synthase; rate of oxygen (O 2 ) consumption; rate of adenosine triphosphate production; mitochondrial DNA content, level of heteroplasmy and any combination thereof 
     
     
         11 . The method of  claim 1 , wherein the administration of the mitochondrially-enriched target cells is by intravenous, intraperitoneal, intraarterial or intramuscular administration. 
     
     
         12 . The method of  claim 1 , wherein between at least 5×10 5  to 5×10 9  mitochondrially-enriched target cells are administered to the subject. 
     
     
         13 . The method of  claim 1 , wherein the mitochondrially-enriched target cells have:
 (a) an increased content of at least one mitochondrial protein selected from SDHA and COX1;   (b) an increased rate of oxygen (O 2 ) consumption;   (c) an increased activity level of citrate synthase;   (d) an increased rate of adenosine triphosphate (ATP) production;   (e) an increased mitochondrial DNA content;   (f) a lower heteroplasmy level; or   (g) any combination thereof,   
       as compared to target cells prior to mitochondrial enrichment. 
     
     
         14 . The method of  claim 1 , further comprising adding a pharmaceutically acceptable carrier to the mitochondrially-enriched target cells prior to administration to the subject. 
     
     
         15 . The method of  claim 1 , wherein the conditions allowing the exogenous mitochondria to enter the target cells comprise incubating the target cells with the exogenous mitochondria at a ratio of about 0.088-176 mU citrate synthase (CS) activity per 10 6  cells. 
     
     
         16 . A pharmaceutical composition for increasing levels of lymphoid cells in a subject comprising mitochondrially-enriched target cells and a pharmaceutically acceptable carrier, wherein the mitochondrially-enriched target cells are enriched with exogenous mitochondria. 
     
     
         17 . The pharmaceutical composition of  claim 16 , wherein the mitochondrially-enriched target cells are produced by the method comprising:
 (a) obtaining target cells from a subject afflicted with a disease or disorder or a donor;   (b) obtaining exogenous mitochondria from a donor; and   (c) producing mitochondrially-enriched target cells by contacting the target cells with the exogenous mitochondria under conditions allowing the exogenous mitochondria to enter the target cells,   
       wherein the mitochondrial content of the mitochondrially-enriched target cells is detectably higher than the mitochondrial content of the target cells. 
     
     
         18 - 30 . (canceled) 
     
     
         31 . A method for diminishing debilitating effects of a lymphocyte deficiency-related disease or diseases in a subject comprising:
 (a) incubating of hematopoietic stem cells (HSCs) with exogenous mitochondria under conditions allowing the exogenous mitochondria to enter the HSCs; and   (b) administering the HSCs from (a) to the subject.   
     
     
         32 . (canceled) 
     
     
         33 . The method of  claim 31 , wherein the exogenous mitochondria have undergone at least one freeze-thaw cycle. 
     
     
         34 . The method of  claim 31 , wherein the conditions allowing the exogenous mitochondria to enter the HSCs comprise a ratio of about 0.088-176 mU citrate synthase (CS) activity per 10 6  cells. 
     
     
         35 - 62 . (canceled)